Abstract The objective of this research is to further develop and clinically validate a real-time multispectral confocal microendoscope for in vivo diagnosis of ovarian cancer. The confocal microendoscope is a new type of instrument for visualizing tissue at the cellular level and has shown great promise for performing optical biopsy. Confocal microendoscopy has the potential to provide a physician with an immediate evaluation of tissue and to survey a much greater area of tissue, reducing the sampling error of traditional tissue extraction biopsy. A confocal microlaparoscope system was constructed and tested in vivo in humans during the prior funding period. A specific aim of this work is to now validate the system and show that it can be used to detect cancer of the ovary during a laparoscopic procedure. Additional objectives of the work are to expand the use of the instrument to the detection of peritoneal implants of ovarian cancer throughout the abdominal cavity and to develop and test a system capable of imaging inside the fallopian tubes. Further technical development is aimed at improving the confocal imaging performance and adding optical coherence tomography as a complimentary imaging modality. The new instrument will incorporate the two imaging modalities into a single compact probe with seamless and rapid switching between modes of operation. In addition to the technology development and clinical translation, another aim of the project is to develop and test targeted contrast agents that provide safer and more effective in vivo identification of ovarian cancer.